The work proposed here seeks to identify small molecules that could be used to validate a novel hypothesis about the origin of inflammation in atherosclerosis and Alzheimer's disease. The hypothesis is that TLR4 together with TLR6 and CD36 respond to the ligands OxLDL and amyloid-? (A?) to promote the inflammation that is at the root of these diseases. Because of the pleiotropic roles for all three of these molecules, standard techniques of gene deletion cannot be used to validate this hypothesis. We will develop enzyme fragment complementation assays using fragments of ?-lactamase fused to TLR4 and TLR6 to enable high throughput screening assays to identify a pool of small molecules with the potential to inhibit TLR4-TLR6 interactions. Counterscreening assays are described to identify which compounds in the pool are specific inhibitors of the binding between TLR4 and TLR6 that do not also inhibit other pairings of TLR4 and TLR6 that are important in contexts other than atherosclerosis and Alzheimer's. These validated compounds will enable further research to explore the hypothesis. If the hypothesis is validated, it will identify a new paradigm for inflammation in atherosclerosis and Alzheimer's disease and could lead to new forms of drug therapy. PUBLIC HEALTH RELEVANCE: Inflammation in atherosclerosis and Alzheimer's disease is a major cause of the pathological consequences of these diseases. This work will explore new ideas about the origin of the inflammation. It will identify new compounds to inhibit the inflammation in model systems and could lead to new drugs for human therapy.